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01.12.2016 | Research | Ausgabe 1/2016 Open Access

Journal of Hematology & Oncology 1/2016

A synthetic three-dimensional niche system facilitates generation of functional hematopoietic cells from human-induced pluripotent stem cells

Journal of Hematology & Oncology > Ausgabe 1/2016
Yulin Xu, Wei Shan, Xia Li, Binsheng Wang, Senquan Liu, Yebo Wang, Yan Long, Ruxiu Tie, Limengmeng Wang, Shuyang Cai, Hao Zhang, Yu Lin, Mingming Zhang, Weiyan Zheng, Yi Luo, Xiaohong Yu, Jiing-Kuan Yee, Junfeng Ji, He Huang
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Electronic supplementary material

The online version of this article (doi:10.​1186/​s13045-016-0326-6) contains supplementary material, which is available to authorized users.



The efficient generation of hematopoietic stem cells (HSCs) from human-induced pluripotent stem cells (iPSCs) holds great promise in personalized transplantation therapies. However, the derivation of functional and transplantable HSCs from iPSCs has had very limited success thus far.


We developed a synthetic 3D hematopoietic niche system comprising nanofibers seeded with bone marrow (BM)-derived stromal cells and growth factors to induce functional hematopoietic cells from human iPSCs in vitro.


Approximately 70 % of human CD34+ hematopoietic cells accompanied with CD43+ progenitor cells could be derived from this 3D induction system. Colony-forming-unit (CFU) assay showed that iPSC-derived CD34+ cells formed all types of hematopoietic colonies including CFU-GEMM. TAL-1 and MIXL1, critical transcription factors associated with hematopoietic development, were expressed during the differentiation process. Furthermore, iPSC-derived hematopoietic cells gave rise to both lymphoid and myeloid lineages in the recipient NOD/SCID mice after transplantation.


Our study underscores the importance of a synthetic 3D niche system for the derivation of transplantable hematopoietic cells from human iPSCs in vitro thereby establishing a foundation towards utilization of human iPSC-derived HSCs for transplantation therapies in the clinic.

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Additional file 1: Growth kinetics of iPSCs seeded in various 3D hematopoietic induction milieu from day 0 to day 10-14. iPSCs were digested with collagenase IV, pipetted gently into small cell aggregations, and then seeded in 3D induction culture. The morphology was captured at indicated time points post seeding. The morphology on day 0 was shown in fig A, D, G, J and M representative of different 3D hematopoietic induction milieu. After 3 days, the formation of a blast colony from iPSC mass was detected as shown in fig B, E, H, K, and N. During induction for 10–14 days, round grape-like cells appeared in 3D induction systems as shown in C, F, I, L, and O. There was no obvious disparity in morphology among various 3D induction systems. (JPG 522 kb)
Additional file 2: Characterization of the committed cells from iPSCs in 3D induction systems. (A) The relative expression of the pluripotent marker OCT4 decreased gradually during the induction phase. (B) The expression of mesoderm transcription factor BRACHYURY was analyzed. (C) Enhanced expression of TAL-1 was detected. (D) The hematopoietic cell markers MIXL1 was significantly elevated during iPSC commitment to CD34+ cells. The values were the mean ± SEM of 3 independent experiments. (E-H) Kinetic expression of NANOG, GATA2, LMO2, and RUNX1 in 3D induction systems using single-cell gene expression analysis. Transcript abundance are calculated with △(35–Ct) values. N = 8. Error bars represented standard error of mean for each sample. (JPG 508 kb)
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